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From 2011.igem.org

July 6, 2011

We did:

1. PCR of the miniprep of Biobrick gen and electrophoresis of the digestion product.

2. PCR of:

-gen Rh1AB, (0,5 microliters)(1)

-gen Rh1AB, (1 microliter)(2)

-Positive control 1 (3)

-Positive control 2 (4)

Rh1AB, (3 microliters)(5)

3. Electrophoresis of the digested parts and undigested with buffer TAE 1x prepared before by Orlando, agarose 1%, 0,1g agar in 100 ml of TAE 1x. Unfortunately electrophoresis did not go. We believe that it can be because the acetic acid concentration was inappropriate.

4. Then we have to do electrophoresis again. This time we did it with TBE 1X Buffer. The agar was made with 0,7g of agarose in 70 ml of TBE 1X recommended because is sure that is sufficient for agarose gel.

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Electrophoresis:

Top side:

- Marker (100-300 K-180)

- Gen uncut

- Gen cut

- Plasmid only Psb1c3

- R+T uncut

- R+T cut

- P+R uncut

- P+R cut

- HindIII marker (125-23130 pb)

Plasmid for parts P+R and R+T is Psb1AK3

Below side:

- Marker (100-300 K-180)

- Negative control

- Positive control 1

- Positive control 2

- Gen 0,5 microliters

- Gen 1 microliter

- Gen 3 microliters

- HindIII marker (125-23130 pb)